CN104604687B - Utilize the method for cutting the induction Multiple Buds propagating quickly butterfly orchid of the bennet stem section after bud - Google Patents
Utilize the method for cutting the induction Multiple Buds propagating quickly butterfly orchid of the bennet stem section after bud Download PDFInfo
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- CN104604687B CN104604687B CN201510060016.6A CN201510060016A CN104604687B CN 104604687 B CN104604687 B CN 104604687B CN 201510060016 A CN201510060016 A CN 201510060016A CN 104604687 B CN104604687 B CN 104604687B
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Abstract
The invention discloses a kind of method of micropropagation of plants, it is to cut the discarded bennet stem section after bud as explant with opening colored iris pedicel axillary buds and induce vegetative bud, directly it is inoculated into the induction that Multiple Buds are carried out in inducing culture, the sprouting induced is cut and is inoculated into progress shoot proliferation culture in proliferated culture medium, the transplanting of strong plantlets and rootage and tissue-cultured seedling is finally carried out.This method made out axillary bud on colored iris bennet and its cut the bennet obsolete material after bud and be able to repeatedly sprout in the culture medium manually prepared to produce sprouting.It the advantage is that neither the influence iris normal florescence is again without injuring iris maternal plant, iris bennet stem section can also be recycled 23 times, make full use of bennet material, turn waste into wealth, substantially increase the proliferative speed and output of iris tissue-cultured seedling, the seedling produced is healthy and strong, neat, is easy to seedling factory management.
Description
Technical field
The induction Multiple Buds of the bennet stem section after bud are cut the invention belongs to iris reproduction technique field, especially a kind of utilize
The method of propagating quickly butterfly orchid.
Background technology
Iris (Phalaenopsis sp.) belongs to orchid family phalaenopsis plant, is found in 1750, most of productions
In moist Asia, NATURAL DISTRIBUTION in A Longmu, Burma, each island in the Indian Ocean, the Malay Peninsula, the South Sea Islands, Philippine with
To the tropical island of the low latitudes such as TaiWan, China.Initial species has kind more than 70, and cenospecies is even more innumerable, its flower color and decorative pattern
Change emerge in an endless stream, spend in vain be, safflower system, pink system, chrysanthemum system, reticulate pattern system, tiger spot system, the kind series such as point line system.
Its flower pattern is peculiar, and attitude is graceful, and pattern is gorgeous, and the florescence is permanent, have the good reputation of " orchid queen ", ornamental value is high, it is deep by
People like that market demand is increasing.
Phalaenopsis seldom develops side shoot in single stem aerial orchid, it is difficult to carry out conventional vegetative propagation；Other iris
Pollination can not be completed under field conditions (factors), seed is hardly resulted in, and embryo is free of in seed, its seed even if carrying out artificial pollination and obtaining
It also is difficult to sprout under breast, natural conditions, germination percentage is extremely low, therefore iris is also difficult by seed and carries out generative propagation.At present
The main quick breeding that iris is carried out using method for tissue culture, belongs to the vegetative propagation of iris, also so-called clonal propagation,
The tissue cultures of iris can obtain a large amount of high-quality, neat iris young plant in a short time, high, fast with proliferation rate
Degree is fast, be not subject to seasonal restrictions, can whole year production and the advantages of easily remove virus, adapt to the wilderness demand in market, be also factory
Change the important channel of nursery.Iris tissue cultures program is generally：(1) explant blade, root, stem apex, bennet etc. are chosen；
(2) induced synthesis protocorm；(3) protocorm is largely bred；(4) plantlet is differentiated；(5) strong plantlets and rootage；(6) greenhouse is transplanted.
It can be seen that the explant that iris tissue cultures are chosen can be blade, root, stem apex, bennet etc. from said procedure (1),
But above-mentioned explant all has certain injury or influence maternal plant normal florescence to maternal plant.The tissue cultures of current iris are normal
There are two kinds of approach, one is that conventional bennet induction vegetative bud is bred, if this method is using existing petal but does not open
Flower or the bennet bloomed can influence iris normal florescence and ornamental value, and breeding coefficient as explant
Each two month is generally 2.0-3.0 or so, and the tissue-cultured seedling produced in the short time is limited；Two are entered using aseptically sowing seeds method
Row iris tissue-cultured seedling is produced, and the seedling variation that this method is bred is more, and seedling early growth is inconsistent.
The content of the invention
The induction Multiple Buds propagating quickly butterfly orchid of the bennet stem section after bud is cut it is an object of the invention to provide a kind of utilize
Method, it is slower to solve iris plant division, is difficult to sprout under seed natural conditions, plant breeding speed is slow, commercial seedling output
Low, growth cycle is inconsistent, flowering time is uneven, it is impossible to high-volume blooming control and listing the problems such as.
To achieve these goals, the present invention is adopted the following technical scheme that, is induced with opening colored iris pedicel axillary buds
Go out vegetative bud and cut the discarded bennet stem section after bud as explant, be directly inoculated into the induction that bud is carried out in inducing culture, will
The sprouting induced, which is cut, is inoculated into proliferated culture medium progress shoot proliferation culture, finally carries out strong plantlets and rootage and tissue-cultured seedling
Transplant.
Specific step is：
(1) selection of explant：The butterfly orchid variety that no disease and pests harm grows fine is chosen, treats that its flower withers, the florescence is
Cross, cut whole piece bennet with scissors, the pedicel axillary buds stem section (one section of one bud) for cutting 3-4cm length is explant material；
(2) preparation of culture medium：
Bud inducement cultivation base：MS+6-BA2.5mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6：
Bud proliferated culture medium：MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6：
Rooting and hardening-off culture base：1/2MS+NAA1.0mg/L+ banana puree 100g/L, sucrose 20g/L, pH value is 5.6；
(3) explant is disinfected：Rinse after 30-60min, above-mentioned explant material in ultra-clean work under running water
In platform, soaking disinfection 45S in 75% alcoholic solution is put into, hereafter alcohol is blotted with aseptic filter paper, 7-10% NaClO is put into
Sterilization 15-20min in thimerosal, then the bract of axillary bud is carefully peelled off, place into 5% on aseptic filter paper
5min is sterilized in NaClO thimerosals, sterilizing uses aseptic water washing 3-5 times after finishing, and filter paper suck dry moisture is placed in standby in culture dish
With；
(4) Fiber differentiation and condition of culture：By the band axillary bud bennet after strict sterilization, with aseptic operation knife by stem section with disappearing
The two ends of venom contact are cut away along 45 DEG C of angles, and bennet bud point enters bud inducement cultivation base with culture base plane into 45 DEG C of angle oblique cuttings upward
In, 25 ± 2 DEG C of cultivation temperature, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is 30-45d left
It is right；
(5) Multiplying culture and condition of culture：The sprouting that step (4) is induced is cut from bennet, and 2-3 plants are one group,
It is inoculated into proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4)：
(6) primary in step (5) is cut into the bennet stem section renewed vaccination after bud into inducing culture, puts culturing rack training
Support, condition of culture and the same step of incubation time (4)；
(7) Multiple Buds that the bennet stem section chain induction after bud goes out will be cut to cut from bennet, 2-3 plants are one group, inoculation
Shoot proliferation, condition of culture and the same step of incubation time (4) are carried out into proliferated culture medium；
(8) repeat step (6) and step (7) 2-3 times；
(9) strong plantlets and rootage and condition of culture：When the sprout that propagation is obtained reaches that 2-5cm is high, under aseptic condition, it will grow thickly
Bud is separated into individual plant, is seeded in Rooting and hardening-off culture base and carries out strong plantlets and rootage, the same step of condition of culture (4), during strong plantlets and rootage
Between for 2-3 months；
(10) training tissue culture seedling and transplanting：When tissue-cultured seedling length is high to 3-5cm, there are 2-5 bar roots, 3-6 piece leaves, the long 2-3cm of leaf
When, you can hardening is carried out, tissue culture bottle is placed on natural lighting condition lower refining seedling 5-7 days, bottle cap is opened and continues hardening 2-3
My god, to strengthen adaptability of the bottle seedling to outdoor environment；It is careful from bottle again to take out tissue-cultured seedling, root culture medium is cleaned, with going out
The sphagna for crossing bacterium is gently wrapped up in the disk of root Zhi Yu36 holes, 25 DEG C or so of keeping temperature, air humidity 80-85%, after 25-30 days,
When thering is the new root young leaves to grow, 1000 times of liquid of carbendazim are periodically sprayed.
Compared to the prior art the present invention the advantage is that：
The invention discloses a kind of using the method that the bennet stem section after bud induces Multiple Buds propagating quickly butterfly orchid is cut, make
Resting bud on the faded iris bennet of flower and its cut the bennet stem section after bud and be able in the culture medium manually prepared
Repeatedly sprout, then bred and culture of rootage, finally obtain complete Phalaenopsis plants.Conventional utilization bennet induction battalion
Support bud to be bred, the growth coefficient in two months is 2.0 or so, and uses as above technical scheme so that each butterfly orchid
The growth coefficient of stalk axillary bud brings up to 14.0-26.0 or so, improves 20 times equivalent to efficiency, has greatly saved and be produced into
This.Advantage of this approach is that the neither influence iris normal florescence is again without injuring iris maternal plant, moreover it is possible to by iris
Bennet stem section is recycled 2-3 times, can be made full use of bennet material, be turned waste into wealth, and substantially increases iris tissue-cultured seedling
Proliferative speed and output, the seedling produced are healthy and strong, neat, are easy to seedling factory management.
Embodiment
With reference to specific embodiment 1, the present invention will be further described.
Embodiment 1：
(1) selection of explant：The black Jack plant of butterfly orchid variety of no disease and pests harm is chosen, treats that flower withers, the florescence is
Cross, cut whole piece bennet with scissors, the pedicel axillary buds stem section (one section of one bud) for cutting 3-4cm length is explant material；
(2) preparation of culture medium：
Bud inducement cultivation base：MS+6-BA2.5mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6：
Bud proliferated culture medium：MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6：
Rooting and hardening-off culture base：1/2MS+NAA1.0mg/L+ banana puree 100g/L, sucrose 20g/L, pH value is 5.6；
(3) explant is disinfected：Rinse after 30-60min, above-mentioned explant material in ultra-clean work under running water
In platform, soaking disinfection 45S in 75% alcoholic solution is put into, hereafter alcohol is blotted with aseptic filter paper, 7-10% NaClO is put into
Sterilization 12-15min in thimerosal, aging material sterilizing 15-20min, it is then on aseptic filter paper that the bract of axillary bud is small
The heart is peelled off, and 5min is sterilized in the NaClO thimerosals for placing into 5%, and sterilizing uses aseptic water washing 3-5 times after finishing, and filter paper is blotted
Moisture is placed in standby in culture dish；
(4) Fiber differentiation and condition of culture：By the band axillary bud bennet after strict sterilization, with aseptic operation knife by stem section with disappearing
The two ends of venom contact are cut away along 45 DEG C of angles, and bennet bud point enters bud inducement cultivation base with culture base plane into 45 DEG C of angle oblique cuttings upward
In, 25 ± 2 DEG C of cultivation temperature, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is 30-45d left
It is right；
(5) Multiplying culture and condition of culture：The sprouting that step (4) is induced is cut from bennet, and 2-3 plants are one group,
It is inoculated into proliferated culture medium and carries out shoot proliferation, culturing room's condition and the same step of incubation time (4)；
(6) primary in step (5) is cut into the bennet stem section renewed vaccination after bud into inducing culture, puts culturing rack training
Support, condition of culture and the same step of incubation time (4)；
(7) Multiple Buds that the bennet stem section chain induction after bud goes out will be cut to cut from bennet, 2-3 plants are one group, inoculation
Shoot proliferation, condition of culture and the same step of incubation time (4) are carried out into proliferated culture medium；
(8) repeat step (6) and step (7) 2-3 times；
(9) strong plantlets and rootage and condition of culture：When the sprout that propagation is obtained reaches that 2-5cm is high, sterile working, by Multiple Buds
Individual plant is separated into, is seeded in Rooting and hardening-off culture base and carries out strong plantlets and rootage, culturing room's conditional synchronization is rapid (4), during strong plantlets and rootage
Between for 2-3 months；
(10) training tissue culture seedling and transplanting：When tissue-cultured seedling length is high to 3-5cm, there are 2-5 bar roots, 3-6 piece leaves, the long 2- of leaf
During 3cm, you can carry out hardening, tissue culture bottle is placed on into natural lighting condition lower refining seedling 5-7 days, bottle cap hardening is opened 2-3 days,
To strengthen adaptability of the bottle seedling to outdoor environment；It is careful from bottle again to take out tissue-cultured seedling, root culture medium is cleaned, with sterilized
Sphagna gently wrap up in the disk of root Zhi Yu36 holes, 25 DEG C or so of keeping temperature, air humidity 80-85% after 30 days, has new root new
When leaf is grown, 1000 times of liquid of carbendazim are sprayed, are periodically sprayed, survival rate to 95%.
It the foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent
With modification, it should all belong to the covering scope of the present invention.
Claims (1)
1. a kind of utilize the method for cutting the induction Multiple Buds propagating quickly butterfly orchid of the bennet stem section after bud, it is characterised in that：With opening
Cross colored iris pedicel axillary buds and induce vegetative bud and cut the discarded bennet stem section after bud as explant, be directly inoculated into induction
The induction of Multiple Buds is carried out in culture medium, the sprouting induced is cut and is inoculated into progress shoot proliferation training in proliferated culture medium
Support, finally carry out the transplanting of strong plantlets and rootage and tissue-cultured seedling；
Specific step is：
(1) selection of explant：The butterfly orchid variety that grows fine of no disease and pests harm is chosen, treats that its flower withers, florescence mistake is used
Scissors cuts whole piece bennet, and the pedicel axillary buds stem section (one section of one bud) for cutting 3-4cm length is explant material；
(2) preparation of culture medium：
Bud inducement cultivation base：MS+6-BA2.5mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6；
Bud proliferated culture medium：MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6；
Rooting and hardening-off culture base：1/2MS+NAA 1.0mg/L+ banana puree 100g/L, sucrose 20g/L, pH value is 5.6；
(3) explant is disinfected：By above-mentioned explant material, rinsed under running water after 30-60min, in superclean bench,
Soaking disinfection 45S in 75% alcoholic solution is put into, hereafter alcohol is blotted with aseptic filter paper, 7-10% NaClO sterilizations are put into
Sterilization 15-20min in liquid, then carefully peels off the bract of axillary bud on aseptic filter paper, and the NaClO for placing into 5% disappears
5min is sterilized in venom, sterilizing uses aseptic water washing 3-5 times after finishing, and filter paper suck dry moisture is placed in standby in culture dish；
(4) Fiber differentiation and condition of culture：By the band axillary bud bennet after strict sterilization, with aseptic operation knife by stem section and thimerosal
The two ends of contact are cut away along 45 DEG C of angles, and bennet bud point enters in bud inducement cultivation base with culture base plane into 45 DEG C of angle oblique cuttings upward,
25 ± 2 DEG C of cultivation temperature, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is 30-45d or so；
(5) Multiplying culture and condition of culture：The sprouting that step (4) is induced is cut from bennet, and 2-3 plants are one group, inoculation
Shoot proliferation, condition of culture and the same step of incubation time (4) are carried out into proliferated culture medium；
(6) primary in step (5) is cut into the bennet stem section renewed vaccination after bud into inducing culture, puts culturing rack culture, trained
The condition of supporting and the same step of incubation time (4)；
(7) Multiple Buds that the bennet stem section chain induction after bud goes out will be cut to cut from bennet, 2-3 plants are one group, are inoculated into increasing
Grow and shoot proliferation is carried out in culture medium, condition of culture and the same step of incubation time (4)；
(8) repeat step (6) and step (7) 2-3 times；
(9) strong plantlets and rootage and condition of culture：When the sprout that propagation is obtained reaches that 2-5cm is high, under aseptic condition, by Multiple Buds point
Strong plantlets and rootage is carried out from into individual plant, being seeded in Rooting and hardening-off culture base, the same step of condition of culture (4), the strong plantlets and rootage time is
2-3 months；
(10) training tissue culture seedling and transplanting：When tissue-cultured seedling length is high to 3-5cm, there are a 2-5 bar roots, 3-6 piece leaves, during the long 2-3cm of leaf, i.e.,
Hardening can be carried out, tissue culture bottle is placed on natural lighting condition lower refining seedling 5-7 days, bottle cap is opened and continues hardening 2-3 days, to increase
Adaptability of the strong bottle seedling to outdoor environment；It is careful from bottle again to take out tissue-cultured seedling, root culture medium is cleaned, with sterilized water
Tongue is gently wrapped up in the disk of root Zhi Yu36 holes, and 25 DEG C or so of keeping temperature, air humidity 80-85% after 25-30 days, has new root new
When leaf is grown, 1000 times of liquid of carbendazim are periodically sprayed.
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Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN108419676A (en) * | 2018-03-30 | 2018-08-21 | 内蒙古自治区生物技术研究院 | Iris tissue culture medium (TCM) and preparation method thereof |
| CN108651444B (en) * | 2018-05-22 | 2020-11-27 | 山东省农业科学院蔬菜花卉研究所 | Preservation and breeding method for phalaenopsis pollen |
| CN109349105A (en) * | 2018-10-10 | 2019-02-19 | 广西生态工程职业技术学院 | A kind of iris tissue-cultured seedling mating system |
| CN109601386B (en) * | 2019-01-24 | 2020-08-25 | 北京农业职业学院 | Method for disinfecting bud cutting explant in stem tip tissue culture of cymbidium sinense |
| CN110896856A (en) * | 2019-11-07 | 2020-03-24 | 郑州师范学院 | Method for inducing complete inflorescence in vitro by axillary bud of inflorescence shaft of phalaenopsis |
| CN111264394A (en) * | 2020-04-07 | 2020-06-12 | 无锡向山兰园科技有限公司 | Tissue culture method of cymbidium hybridum |
| CN111758567B (en) * | 2020-05-29 | 2022-07-12 | 福建农林大学 | Phalaenopsis single-bud propagation method adopting symbiotic microbial inoculum |
| CN113197098A (en) * | 2021-06-07 | 2021-08-03 | 芜湖东源新农村开发股份有限公司 | Inoculation method for shortening rapid propagation growth cycle of phalaenopsis |
| CN113575421A (en) * | 2021-08-27 | 2021-11-02 | 中德润萌生态科技(青岛)有限公司 | Aseptic processing method for tissue culture material of orchidaceae plant |
| CN113973713A (en) * | 2021-10-28 | 2022-01-28 | 北京市昌平职业学校 | Butterfly orchid tissue culture method |
| CN115251057B (en) * | 2022-08-26 | 2023-07-25 | 郑州师范学院 | Method for inducing germination of phalaenopsis seedlings by using plant hormone composition |
| CN116686715A (en) * | 2023-07-25 | 2023-09-05 | 云南省农业科学院花卉研究所 | Butterfly orchid tissue culture seedling cultivation method without carrying cymMV and ORSV |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH09271282A (en) * | 1996-04-03 | 1997-10-21 | Sapporo Breweries Ltd | Manufacture of moth orchid seedling by root apex cultivation |
| CN1258321C (en) * | 2004-07-30 | 2006-06-07 | 中国科学院华南植物园 | Quick breeding method for butterfly orchid high quality sprout |
| CN1994068A (en) * | 2006-12-27 | 2007-07-11 | 李�杰 | Method for quickly breeding butterfly orchid by using pedicel nodal bud to induct pedicel |
| CN102613076A (en) * | 2012-03-26 | 2012-08-01 | 吴海红 | Vegetative propagation method for butterfly orchid |
| CN103766218B (en) * | 2013-12-27 | 2015-05-13 | 佛山市顺德区今日景艺生物科技有限公司 | Butterfly orchid induction culture medium and asexual propagation method of butterfly orchid |
| CN104082148B (en) * | 2014-07-18 | 2016-08-17 | 内蒙古农业大学 | The method of iris aseptic bennet regeneration expanding propagation |
| CN104067943B (en) * | 2014-07-18 | 2017-02-15 | 内蒙古农业大学 | Phalaenopsis sterile root propagation method |
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